Document Detail

Ultra-low-volume, real-time measurements of lactate from the single heart cell using microsystems technology.
MedLine Citation:
PMID:  11866072     Owner:  NLM     Status:  MEDLINE    
The fabrication of microelectrodes integrated within ultra-low-volume microtiter chambers for the amperometric determination of metabolites continues to be of interest in the subject of single-cell and high-throughput screening. The microsystem described in this paper consists of a two-microelectrode sensor with a microfluidic dispensation technology, which is able to deliver both very low titers (6.5 pL) and single heart cells into a low-volume microphotoelectrochemical cell. Devices were fabricated using photolithography and liftoff giving reproducible sensors integrated within high aspect ratio titer chambers (with a volume of 360 pL), made of the photoepoxy SU8. In this paper, the determination of lactate was optimized using an enzyme-linked assay based upon lactate oxidase, involving the amperometric determination of hydrogen peroxide at +640 mV versus an internal Ag/AgCl pseudoreference. The microsystem (including the microfluidic dispensers and structures as well as the microsensor) was subsequently used to measure the lactate content of single heart cells. Dynamic electrochemical measurements of lactate during cell permeabilization are presented. We also show the use of respiratory uncouplers to simulate ischemia in the single myocyte and show that, as expected, the rate of lactate production from the hypoxic heart cell is greater than that within the normoxic healthy myocyte.
Xinxia Cai; Norbert Klauke; Andrew Glidle; Peter Cobbold; Godfrey L Smith; Jonathan M Cooper
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  74     ISSN:  0003-2700     ISO Abbreviation:  Anal. Chem.     Publication Date:  2002 Feb 
Date Detail:
Created Date:  2002-02-27     Completed Date:  2002-04-01     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  908-14     Citation Subset:  IM    
Bioelectronics Research Centre, Department of Electronics and Electrical Engineering, University of Glasgow, UK.
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MeSH Terms
Indicators and Reagents
Lactic Acid / analysis*
Lipoxygenase / chemistry
Metals / chemistry
Myocardium / chemistry*,  cytology*
Reg. No./Substance:
0/Indicators and Reagents; 0/Metals; 50-21-5/Lactic Acid; EC

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